博碩士論文 972204021 詳細資訊




以作者查詢圖書館館藏 以作者查詢臺灣博碩士 以作者查詢全國書目 勘誤回報 、線上人數:20 、訪客IP:3.226.245.48
姓名 張瑋仁(Wei-Jen Chang)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 血清素受體2B參與血清素引起的機械性痛覺過敏
(Serotonin receptor 2B is involved in 5-HT-induced mechanical hyperalgesia)
相關論文
★ 週邊發炎反應增加酸敏感受體- TDAG8基因在背根神經節之表現量★ 酸敏感G蛋白偶合受體,G2A,在ASIC3基因剔除小鼠中改變表現量
★ MrgB4受體專一表現於感覺神經元,且在ASIC3基因剔除小鼠中有不同的表現。★ 血清素受體2B對酸敏感離子通道3與辣椒素受體1的影響
★ 酸敏感G蛋白偶合受體在小鼠背根神經節神經元中的訊息傳導路徑★ 酸敏感G蛋白偶合受體功能上的拮抗機制
★ TDAG8活化後經由PKA與PKCε增強辣椒素受體的敏感度★ 台灣海岸植物之內生真菌多樣性研究
★ ASIC3、TRPV1或TDAG8基因缺失會減緩關節炎誘導的熱痛覺過敏並抑制衛星膠細胞表現★ 抑制OGR1表現可減緩慢性神經性疼痛藉由減少顆粒性白血球數及非IB4神經元之鈣訊號
★ 抑制OGR1及G2A表現可藉由調控非IB4神經元鈣訊號減緩酸所誘導長期疼痛★ G-蛋白偶合接受體與G-蛋白訊號調控蛋白之整合型資料庫
★ 血清素受體2B基因在酸敏感受體3基因剔除小鼠的背根神經節中表現量增加★ 酸敏感的G蛋白偶合受體─OGR1表現在背根神經節內與痛覺相關的感覺神經元上
★ 血清素受體2B調控鈣離子變化影響機械性痛覺敏感★ 蛋白質激酶A以及蛋白質激酶Cɛ在急性轉換至慢性發炎性疼痛中扮演的角色
檔案 [Endnote RIS 格式]    [Bibtex 格式]    [相關文章]   [文章引用]   [完整記錄]   [館藏目錄]   [檢視]  [下載]
  1. 本電子論文使用權限為同意立即開放。
  2. 已達開放權限電子全文僅授權使用者為學術研究之目的,進行個人非營利性質之檢索、閱讀、列印。
  3. 請遵守中華民國著作權法之相關規定,切勿任意重製、散佈、改作、轉貼、播送,以免觸法。

摘要(中) 血清素 (Serotonin)是發炎物質的一種,並且主要表現在中樞和週邊神經系統。在受傷或是發炎後,血清素會從血小板或是肥大細胞釋放到週邊神經系統,並引起痛覺或是痛覺過敏。血清素受體共有七個亞型。然而目前還不清楚哪一個血清素受體和血清素引起的痛和痛覺過敏相關。在這篇論文裡,我研究血清素受體2B在血清素引起的痛和痛覺過敏中所扮演的角色。我發現注射血清素或是血清素2的促進劑會產生對機械性刺激的痛覺過敏。而血清素引起的機械性痛覺過敏可以被血清素受體2B/2C的抑制劑給抑制,但血清素受體2A的抑制劑不能抑制血清素引起的痛覺過敏,意指血清素受體2B或2C參與了血清素引起痛覺過敏。考慮到血清素受體2C沒有表現在背根神經節中,血清素受體2B參與血清素引起的機械性痛覺過敏中。進一步我想了解血清素受體2B調控機械性痛覺過敏的機制,我共同轉染了血清素受體2B和與痛覺過敏相關的離子通道(例如酸敏感離子通道家族或辣椒素受體1)到人類胚胎腎臟細胞中,並且以鈣離子攝影驗證血清素受體2B是否會調控這些離子通道。
摘要(英) Serotonin (5-hydroxtryptamine [5-HT]) is one of the inflammatory mediators present in central and peripheral nervous system. After injury or inflammation, 5-HT is released from platelets and mast cells to peripheral nervous system, causing pain and hyperalgesia. 5-HT receptors include seven subtypes (5HT1-7). However, it remains unclear which subtype of 5-HT receptors is involved in 5-HT-induced pain and hyperalgesia. In this study, we have investigated the roles of 5-HT2B receptors in 5-HT-induced pain and hyperalgesia. We have found that injection of 5-HT or 5-HT2 agonist ?-m-5HT produces significant hyperalgesia to mechanical stimuli. The mechanical hyperalgesia induced by 5-HT injection is inhibited by 5-HT2B/2C antagonist SB206553, but not 5-HT2A antagonist Ketanserin, suggesting that 5-HT2B or 5-HT2C receptors are involved in 5-HT-induced mechanical hyperalgesia. Given that 5-HT2C receptor is not present in not present in DRG, it could be 5-HT2B receptor involved in 5-HT-induced mechanical hyperalgesia. To further understand the mechanism of 5HT2B-mediated mechanical hyperalgesia, we co-transfect 5-HT2B and ion channels that are related to hyperalgesia (such as ASICs or TRPV1) to human embryo kidney 293T cells (HEK293T), and examine whether 5-HT2B receptor regulates these ion channels by calcium image.
關鍵字(中) ★ 血清素
★ 血清素受體2B
★ 機械性痛覺過敏
關鍵字(英) ★ hyperalgesia
★ serotonin
★ 5-HT
★ 5-HT2B
論文目次 摘要 i
Abstract ii
致謝 iii
目錄 iv
圖目錄 vii
縮寫與全名對照表 viii
第一章 緒論 1
1.1 痛覺 2
1.2痛覺的訊息傳導途徑 2
1.3 發炎物質引起的痛覺過敏 3
1.3.1 血清素引起的痛覺過敏 3
1.4血清素受體的分類與路徑 4
1.5與痛覺敏感相關的血清素受體 4
1.5.1血清素受體1A 4
1.5.2血清素受體2A 5
1.5.3血清素受體3A 5
1.5.4血清素受體4 5
1.5.5血清素受體7 6
1.6與機械性痛覺過敏相關的離子通道 6
1.6.1酸敏感離子通道家族 (acid-sensing ion channels, ASICs) 6
1.6.2辣椒素受體1 (transient receptor potential cation channel, subfamily V, member 1, TRPV1) 7
1.6.3電壓調控鈉離子通道 (Voltage-gated sodium channels, Nav ) 7
1.7研究動機與目的 8
第二章 材料與方法 9
2.1實驗材料 10
2.1.1 菌株 10
2.1.2 細胞株 10
2.1.3 實驗動物 10
2.1.4 藥品 10
2.2 實驗方法 11
2.2.1 膠體萃取法 (gel extraction) 11
2.2.2瓊脂膠體的製備及電泳 11
2.2.3聚合酵素連鎖反應 (Polymerase chain reaction, PCR) 11
2.2.4 大腸桿菌的轉型作用(Transformation) 12
2.2.5 細菌培養 12
2.2.6 質體製備 13
2.2.7 細胞培養 14
2.2.8轉染作用(Transfect) 14
2.2.9 鈣離子分析(Calcium imaging) 15
2.2.10動物疼痛行為實驗 16
2.2.11 統計分析 17
第三章 結果 18
3.1 血清素受體2B會受到血清素或血清素受體2促進劑的活化,並且被血清素受體2B/2C的抑制劑抑制 19
3.2 血清素受體2A會受到血清素及血清素受體2促進劑的活化,並且被血清素受體2A的抑制劑抑制 19
3.3 血清素受體2C會受到血清素的活化,並且被血清素受體2B/2C的抑制劑抑制 20
3.4 血清素受體1A會受到血清素的活化,並且被血清素受體1A的抑制劑抑制 20
3.5 血清素受體3A會受到血清素的活化,並且被血清素受體3A的抑制劑抑制 20
3.6 血清素受體2B/2C的抑制劑可以抑制由血清素所引發的機械性痛覺異常,但不能抑制血清素引起的腳腫脹 21
3.7血清素受體2B/2C的抑制劑可以抑制由血清素受體2促進劑所引發的機械性痛覺異常 21
3.8 抑制血清素受體1A和3A的活性不能抑制由血清素所引發的機械性痛覺過敏 22
3.9血清素受體2B不會抑制由血清素引起的熱痛覺過敏 23
3.10 血清素受體2B/2C的抑制劑不能抑制由發炎物質所引發的機械性痛覺過敏 23
3.11 血清素受體2B/2C的抑制劑不能抑制由辣椒素和血清素加pH5.0生理緩衝混合液所引發的機械性痛覺過敏 24
3.12 抑制鈉離子通道可以抑制機械性痛覺過敏的產生 24
3.13 轉染辣椒素受體1,酸敏感離子通道1和3至人類胚胎腎臟細胞大量表現後,受不同濃度辣椒素和不同pH值的酸性溶液刺激後所產生的反應 25
3.14在共同轉染血清素受體2B和辣椒素受體1的細胞中,血清素受體2B的活化會增強辣椒素受體1對鈣離子通透性 25
3.15在共同轉染血清素受體2B和酸敏感離子通道1a和3的細胞中,血清素受體2B的活化會減少酸敏感受體1a和3對鈣離子通透性 26
第四章 討論 28
4.1血清素受體2B參與血清素引起的機械性痛覺過敏 29
4.2血清素受體2B為主要調控血清素引起的痛覺過敏 29
4.3 血清素受體3引起短暫的疼痛,並影響痛覺過敏的維持 30
4.4血清素受體1A對血清素引起的機械性痛覺過敏有些微抵抗疼痛的效果 31
4.5 血清素受體2B/2C不參與辣椒素和血清素加pH5.0酸性生理緩衝液所引發的機械性痛覺過敏 31
4.6血清素受體2B/2C不會調控由發炎物質所引起的機械性痛覺過敏 31
4.7抑制鈉離子通道可抑制機械性痛覺過敏的產生 32
4.8血清素受體引發痛覺過敏的機制 32
第五章 參考文獻 34
附錄 72
圖目錄
圖3.1 血清素受體1A、 2A、 2B、 2C、 3A和4基因圖譜 41
圖3.2 血清素受體2B受到促進劑和抑制劑刺激後調控細胞內鈣離子濃度 43
圖3.3 血清素受體2A受到促進劑和抑制劑刺激後調控細胞內鈣離子濃度 45
圖3.4 血清素受體2C受到促進劑和抑制劑刺激後調控細胞內鈣離子濃度 47
圖3.5 血清素受體1A受到促進劑和抑制劑刺激後調控細胞內鈣離子濃度 49
圖3.6 血清素受體3A受到促進劑和抑制劑刺激後調控細胞內鈣離子濃度 50
圖3.7 血清素受體2B/2C的抑制劑可以抑制由血清素所引發的機械性痛覺異常,但血清素受體2A的抑制劑不能 52
圖3.8 血清素受體2B/2C的抑制劑可以抑制由血清素2促進劑所引發的機械性痛覺異常,但血清素受體2A的抑制劑不能 54
圖3.9 血清素受體1A和3A的抑制劑不能抑制由血清素所引發的機械性痛覺異 56
圖3.10 血清素受體2A的抑制劑可以抑制由血清素所引發的熱痛覺過敏 58
圖3.11 血清素受體2B/2C的抑制劑不能抑制由CFA和carrageenan所引發的機械性痛覺異常 60
圖3.12 血清素受體2B/2C的抑制劑不能抑制由血清素與酸/辣椒素所引發的機械性痛覺異常 62
圖3.13 鈉離子通道的抑制劑能抑制由血清素所引發的機械性痛覺異常 64
圖3.14在人類胚胎腎臟細胞中各種基因的分布 65
圖 3.15 在酸敏感離子受體1a、3和辣椒素受體1基因大量表現的人類腎臟胚胎細胞中,鈣離子變化量和血清素濃度的關係圖 66
圖3.16 血清素受體2B能增強辣椒素受體1的功能 69
圖3.17 血清素受體2B抑制酸敏感離子通道1a和3的功能 71
參考文獻 Amaya, F., Wang, H., Costigan, M, Allchorne, A. J., Hatcher, J. P., Egerton, J., Stean, T., Morisset, V., Grose, D., Gunthorpe, M. J., Chessell, I. P., Tate, S., Green, P. J., Woolf, C. J., (2006). "The voltage-gated sodium Channel Nav1.9 is an effector of peripheral inflammatory pain hypersensitivity" J. Neuroscience 26(50): 12852-12860.
Brenchat, A., Romero, L., Garcia, M., Pujol, M., Burgueno, J., Torrens, A., Hamon, M., Baeyens, J., Buschmann, H., Zamanillo, D. (2009)" 5-HT7 receptor activation inhibits mechanical hypersensitivity secondary to capsaicin sensitization in mice." Pain 141(3): 239-247.
Caterina, M. J., Schumacher, M.A., Tominaga, M., Rosen, T.A., Levine, J.D., Julius, D. (1997) "The capsaicin receptor: a heat-activated ion channel in the pain pathway" Nature 389(6653): 816-824.
Cardenas, L. M., Cardenas, C.G., Scroggs, R.S. (2001). "5HT Increases excitability of nociceptor-like rat dorsal root ganglion neurons via cAMP-coupled TTX-resistant Na+ Channels" J. Neurophysiol 86: 241-248.
Chen, C. C., England, S., Akopian, A.N., Wood, J.N. (1998). "A sensory neuron-specific, proton-gated ion channel" Proc. Natl. Acad. Sci. USA 95: 10240-10245.
Corey, D. P., Garcia-Anoveros, J. (1996). "Mechanosensation and the DEG/ENaC Ion Channels" Science 273(5273): 361-364.
Davis, J. B., Gray, J., Gunthorpe, M.J., Hatcher, J.P., Davey, P.T., Overend, P., Harries, M.H., Latcham, J., Clapham, C., Atkinson, K., Hughes, S.A., Rance, K., Grau, E., Harper, A.J., Pugh, P.L., Rogers, D.C., Bingham, S., Randall, A., Sheardown, S.A. (2000). "Vanilloid receptor-1 is essential for inflammatory thermal hyperalgesia" Nature 405(6783): 183-187.
Doak, G. J., Sawynok, J. (1997). "Formalin-induced nociceptive behavior and edema: involvement of multiple peripheral 5-hydroxytryptamine receptor subtypes " Neuroscience 80(3): 939-949.
Dray, A. (1995). "Inflammatory mediators of pain" British Journal of Anaesthesia 75: 125-131.
Giordano, J., Dyche, J. (1989). "Differential analgesic actions of serotonin 5-HT3 receptor antagonists in the mouse" Neuropharmacology 29(4): 423-427.
Goldin, A. L., Barchi, R.L. Caldwell, J.H., Hofmann, F., Howe, J.R., Hunter, J.C., Kallen, R.C. Mandel, G. Meisler, M.H. Netter, Y.B. Noda, Y. Tamkun, M.M. Waxman, S.G. Wood, J.N. Catterall, W.A. (2000). "Nomenclature of voltage-gated sodium channels" Neuron 28(2): 365-368.
Granados-Soto, V., Argüelles, C. F., Rocha-González, H. I., Godínez-Chaparro, B., Flores-Murrieta, F. J., Villalón, C. M. (2010) "The role of peripheral 5-HT1A, 5-HT1B, 5-HT1D, 5-HT1E and 5-HT1F serotonergic receptors in the reduction of nociception in rats" Neuroscience 165(2): 561-568.
Hamamotoa, D. T. O.-G. l., X. R., Hondaa, J. M., Kajander K. C. (1998). "Intraplantar injection of hyaluronic acid at low pH into the rat hindpaw produces tissue acidosis and enhances withdrawal responses to mechanical stimuli." Pain 74: 225-234.
Huang and Sun (2005). “Enhanced transcription of serotonin receptor 2B in dorsal root ganglion from ASIC3 knockout mice” master thesis. Department of Life Sciences. Jhongli, National Central University.
Jarvis, M. F., Honore, P., Shieh, C. C., Chapman, M., Joshi, S., Zhang, X. F., Kort, M., Carroll, W., Marron, B., Atkinson, R., Thomas, Liu, D., Krambis, M., Liu, Y., McGaraughty, S., Chu, K., Roeloffs, R., Zhong, C., Mikusa, J. P., Hernandez, G., Gauvin, D., Wade, C., Zhu, C., Pai, M., Scanio, M., Shi, L., Drizin, I., Gregg, R., Matulenko, M., Hakeem, A., Gross, M., Johnson, M.,Marsh, K., Wagoner, P. K., Sullivan, J. P., Faltynek, C. R., Krafte, D. S. (2007). "From the Cover: A-803467, a potent and selective Nav1.8 sodium channel blocker, attenuates neuropathic and inflammatory pain in the rat" Proceedings of the National Academy of Sciences 104(20): 8520-8525.
Julius, D., basbaum, A. (2001). "Molecular mechanisms of nociception." Nature 413: 203-210.
Kayser, V., Elfassi, I., Aubel, B., Melfort, M., Julius, D., Gingrich, J., Hamon, M., Bourgoin, S. (2007). "Mechanical, thermal and formalin-induced nociception is differentially altered in 5-HT1A−/−, 5-HT1B−/−, 5-HT2A−/−, 5-HT3A−/− and 5-HTT−/− knock-out male mice" Pain 130(3): 235-248.
Lingueglia, E. (2007). "Acid-sensing Ion channels in sensory perception" JBC Papers in Press 282(24): 17325-17329.
Lin and Sun (2007). “The effects of serotonin receptor 2B on the acid-sensing ion channel 3 (ASIC3) and transient receptor potential cation channel 1(TRPV1)” master thesis. Department of Life Sciences. Jhongli, National Central University.
Luz M. Cardenas, C. G. C., and Reese S. Scroggs (2001). "5-HT increases excitability of nociceptor-like rat dorsal root ganglion neurons via cAMP-coupled TTX-resistant Na+ channels" J Neurophysiol 86: 241-248.
Marban, E., Yamagishi, T., Tomaselli, G. F. (1998). "Structure and function of voltage-gated sodium channels" Journal of Physiology 508(3): 647-657.
McGowan, E., Hoyt, S.B. Li, X., Lyons, K.A., Abbadie, C. (2009). "A peripherally acting Nav1.7 sodium channel blocker reverses hyperalgesia and allodynia on rat models of inflammatory and neuropathic pain" Anesthesia&Analgesia 109(3): 951-958.
Millan, M. J. (2002). "Descending control of pain" Progress in Neurobiology 66: 355–474.
Mohammad-Zadeh, L. F., Moses, L., Gwaltney-Brant, S. M. (2008). "Serotonin: a review" Journal of Veterinary Pharmacology and Therapeutics 31(3): 187-199.
Neer, E. J. (1995). "Heterotrimeric C proteins: Organizers of transmembrane signals " Cell 80(2): 249-257.
Nichols, D. E., Nichols, C.D. (2008). "Serotonin Receptors" Chemical Reviews 108: 1614-1641.
Nicholson, R., Small, J.,Dixonc, A.K.,Spanswickb, D., Lee, K. (2003). "Serotonin receptor mRNA expression in rat dorsal root ganglion neurons" Neuroscience Letters 337(3): 119-122.
Ohta, T., Y. Ikemi, et al. (2006). "Potentiation of transient receptor potential V1 functions by the activation of metabotropic 5-HT receptors in rat primary sensory neurons." The Journal of Physiology 576(3): 809-822.
Price, M. P., McIlwrath, S. L., Xie, J., Cheng, C., Qiao, J., Tarr, D.E., Sluka, K.A., Brennan, T.J., Lewin, G.R,, Welsh, M.J. (2001). "The DRASIC Cation Channel Contributes to the Detection of Cutaneous Touch and Acid Stimuli in Mice" Neuron 32(1071-1083): 1071.
Schmelz, M., Schmidt R.,Weidner C. , Hilliges, M.,Torebj¨ork, H. E., Handwerker, H. O. (2003). "Chemical Response Pattern of Different Classes of C-Nociceptors to Pruritogens and Algogens" Journal of Neurophysiology 89(5): 2441-2448.
Scholz, J. and C. J. Woolf (2002) "Can we conquer pain?" Nature Neuroscience 5(Supp): 1062-1067.
Staniland, A. A., McMahon, S. B. (2009). "Mice lacking acid-sensing ion channels (ASIC) 1 or 2, but not ASIC3, show increased pain behaviour in the formalin test" European Journal of Pain 13(6): 554-563.
Sufka, K. J., Schomburg F. M. and Giordano J. (1991). "Receptor mediation of 5-HT-Induced inflammation and nociception in Rats" Pharmacology Biochemistry & Behavior 41: 53-56.
Taiwo, Y. O., Levine, J. D. (1992). "Serotonin is a directly-acting hyperalgesic agent in the rat" Neuroscience 48(2): 485-490.
Tokunaga, A., Saika, M., Senba, E. (1998). "5-HT2A receptor subtype is involved in the thermal hyperalgesic mechanism of serotonin in the periphery" Pain 76: 349-355.
Treede, R.-D. (2003). "Neurophysiological studies of pain pathways in peripheral and central nervous system disorders" Journal of Neurology 250(10): 1152-1161.
Waldmann, R., Champigny, G., Bassilana, F., Heurteaux, C., Lazdunski, M. (1997). "A proton-gated cation channel involved in acid-sensing" Nature 386: 173-177.
Waldmann, R., Bassilana, F., Weille, J. D., Champigny, G., Heurteaux, C., Lazdunski, M. (1997). "Molecular Cloning of a Non-inactivating Proton-gated Na1 Channel Specific for Sensory Neurons" The Journal of Biological Chemistry 272(34): 20975-20978.
Waldmann, R., Lazdunski, M. (1998). "H+-gated cation channels: neuronal acid sensors in the NaC/DEG family of ion channels " Current Opinion in Neurobiology 8(3): 418-424.
Walker, K. M., Urban L., Medhurst, S. J., Patel, S., Panesar, M., Fox, A. J., McIntyre, P. (2003). "The VR1 Antagonist Capsazepine Reverses Mechanical Hyperalgesia in Models of Inflammatory and Neuropathic Pain" Journal of Pharmacology and Experimental Therapeutics 304(1): 56-62.
Wu, S. X., Zhua, M., Wanga, W., Wanga, Y.Y. Li, Y.Q.,Yew, D.T. (2001). "Changes of the expression of 5-HT receptor subtype mRNAs in rat dorsal root ganglion by complete Freund's adjuvant-induced infammation" Neuroscience Letters 307: 183-186.
Voilley, N., Weille, J.D., Mamet, J., Lazdunski, M. (2001). "Nonsteroid Anti-Inflammatory Drugs Inhibit Both the Activity and the Inflammation-Induced Expression of Acid-Sensing Ion Channels in Nociceptors "The Journal of Neuroscience 21(20):8026–8033.
Zeitz, K. P., Guy, N., Malmberg, A. B. Dirajlal, S., Martin, W. J. Sun, L., Bonhaus, D. W., Stucky, C.L. Julius, D., Basbaum, A. I. (2002). "The 5-HT3 Subtype of Serotonin Receptor Contributes to Nociceptive Processing via a Novel Subset of Myelinated and Unmyelinated Nociceptors" The Journal of Neuroscience 22(3): 1010-1019.
指導教授 孫維欣(Wei-Hsin Sun) 審核日期 2010-10-22
推文 facebook   plurk   twitter   funp   google   live   udn   HD   myshare   reddit   netvibes   friend   youpush   delicious   baidu   
網路書籤 Google bookmarks   del.icio.us   hemidemi   myshare   

若有論文相關問題,請聯絡國立中央大學圖書館推廣服務組 TEL:(03)422-7151轉57407,或E-mail聯絡  - 隱私權政策聲明